Expression profiles and differential diagnostic value of serum Golgi protein-73 in patients with liver cirrhosis and primary hepatic carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression profiles of serum Golgi protein-73 (GP73) in liver cirrhosis and primary hepatic carcinoma (PHC) and determine its clinical value for differential diagnosis.</p><p><b>METHODS</b>Serum protein expressions of GP73 and alpha-fetoprotein (AFP) were detected by enzyme-linked immunosorbent assay and chemiluminescence assay, respectively, in patients with PHC (n=80), liver cirrhosis (n=65), and healthy controls (n=50). Inter-group changes were assessed by Kruskal-Wallis test, and significance of these differences was assessed by Mann-Whitney test. A receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency and determine the cut-off values for GP73 and AFP. Sensitivity and specificity were compared by the Chi-squared test. Correlation between serum GP73 expression and clinical parameters was determined by Spearman's rank correlation analysis.</p><p><b>RESULTS</b>The PHC group showed significantly higher serum GP73 (282.0 mug/L) than the liver cirrhosis group (211.8 mug/L) and control group (58.3 mug/L) (H = 93.30, P less than 0.01). For differential diagnosis of PHC and liver cirrhosis, the cut-off value was 318.1 mug/L for GP73 and 13.4 mug/L for AFP. Sensitivity of GP73 was lower than AFP (45% (36/80) vs. 65% (52/80); X2 = 8.02, P less than 0.05). Specificity of GP73 was lower than AFP but no significance was found (83.1% (54/65) vs. 87.7% (57/65); X2=0.27, P more than 0.05). The areas under the ROC curves were not significantly different between GP73 and AFP (0.65 (95% confidence interval (CI): 0.54~0.72) vs. 0.75 (95% CI: 0.67~0.83); Z = 1.88, P more than 0.05). The area under the ROC curves increased but not significantly (0.80 (95% CI: 0.73~0.88) vs. 0.75 (95% CI: 0.67~0.83); Z=2.61, P more than 0.05). Serum GP73 was correlated with liver cirrhosis (r=0.27), vascular invasion (r=0.29), and TNM staging (r=0.27) (all P less than 0.05), but not with sex (r=0.13), age (r=0.10), enhanced AFP (> 13.4 mug/L; r=0.03), tumor size (r=0.18), or distant metastasis (r=0.04), all P less than 0.05.</p><p><b>CONCLUSION</b>Serum GP73 and AFP have comparable diagnostic efficiency, but the sensitivity of AFP is superior for differential diagnosis of liver cirrhosis and primary hepatic carcinoma. Elevated serum GP73 may be correlated with liver tumor load and aggressiveness.</p>

Secretory expression and biological activity analysis of an anti-H5 single-chain antibody from Pichia pastoris / 病毒学报

In our previous study, a panel of 52 broadly cross-reactive H5-specific monoclonal antibodies (MAbs) were generated and characterized. The 13D4, one of these MAbs, has been demonstrated to protect mice against lethal challenge by 4 strains of H5N1 avian influenza virus representing the currently prevailing genetic populations, clades 1, 2.1, 2.2, and 2.3. Here, we further cloned the gene of the 13D4 MAb and constructed a single-chain variable fragment. Then, the 13D4 single-chain antibody (scFv) was expressed in secretory maner in Pichia pastoris. The supernatant of the culture was concentrated and subjected to ammonium sulfate precipitation. The purity of the 13D4 scFv was around 90% in SDS-PAGE following ion-exchange chromatography. We further investigated its binding property using hemagglutination inhibition (HI) test and blocking ELISA. The results indicated that the 13D4 scFv shared the same binding sites and comparable HI titer with the prototype murine 13D4 Mab. In conclusion, an anti-H5 single-chain wide-spectrum neutralizing antibody is prepared successfully in yeast system.

Establishment of serological detection methods of 3 types of human herpesvirus 8 specific-antigens and complex antigens / 中华传染病杂志

Objective To establish serological detection methods of human herpesvirus 8 (HHV-8).Methnds Three potent antigenic fusion proteins.K8.1,ORF65 and ORF73 C of HHV- 8 were synthesized using E.coli system.The sera were detected using lhese antigenic proteins.The positive sera were from 12 patients with Kaposi's sarcoma and 32 patients with acquired immunodeficiency syndrome-related Kaposi's sarcoma.The negative sera were from 20 patients with cutaneous tumors and children under 15 years old.Western blot and enzyme-linked immunosorbent assay (EI.ISA)were employed to determine the immunogenicity of each recombinant protein and the sensitivity and specificity of ELISA using the complex antigens.Results Three types highly purified HHV 8 specific recombinant pro teins with potent antigenicity were successfully synthesized.The sensitivity of ELISA using the above complex antigens was significantly higher than traditional immuno-flurescent assay (IFA)detecting the positive and negative sera,whieh were 81.8%,34.4%,respectively.And the specificity of ELISA was 97.9%.Conclusion K8.1,ORF65 and ORE73 C are good candidate antigens for establishing HHV-8 serological detection methods,which have better sensitivity and specificity.